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Histological and Morphometric Analysis of Testes of Adult Quail (Coturnix Coturnix Japonica) of Bangladesh

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Histological and Morphometric Analysis of Testes of Adult Quail (Coturnix Coturnix Japonica) of Bangladesh Int. J. Morphol., 33(1):100-104, 2015. Histological and Morphometric Analysis of Testes of Adult Quail (Coturnix coturnix japonica) of Bangladesh Análisis Histológico y Morfométrico de Testiculos de Codorniz Adulta (Coturnix coturnix japonica) de Bangladesh Subrata Kumar Shil*; Md. Abul Quasem* & Mohammad Lutfur Rahman* SHIL, S. K.; QUASEM, A. & RAHMAN, M. L. Histological and morphometric analyses of testes of adult quail (Coturnix coturnix japonica) of Bangladesh. Int. J. Morphol., 33(1):100-104, 2015. SUMMARY: A histological and morphometric study was conducted to evaluate the seasonal testicular variations in the adult Japanese quail (Coturnix coturnix japonica) in Bangladesh. The weight of birds and testes, volume, length, circumference, seminiferous tubule diameter, circumference and height of germinal epithelium of testes were measured. Sections of testes obtained from studied birds were stained with H&E and images were taken by J software. Weight and volume of paired testes of quails significantly heavier (P<0.05) in long day period of summer and rainy season (March – September) than the short day period of autumn and winter (October – February). Seasons had no significant effect on live weight gain of quails. The weight and diameter of testes or height of the germinal epithelium of the seminiferous tubule were positively correlated (r=0.84; r=0.88). Spermatogenic activity was pronounced during summer and rainy season than that of autumn and winter (P<0.05). KEY WORDS: Japanese quail; Morphometry; Testis; Season; Day length; Temperature. INTRODUCTION Reproduction of birds depends on photoperiod, in temperate zone species (Wingfield et al., 1992; Ball, rainfall, humidity, temperature and food as well as intrinsic 1993; Cockrem, 1995; Hahn et al., 1997). Bangladesh has factors (Lake, 1981; Lofts & Murton, 1968). In temperate a subtropical monsoon climate characterized by wide zones, seasonal day length variation serves as the primary seasonal variations in rainfall, moderately warm role of timing reproductive periodicity (Lofts & Murton). temperatures, and high humidity and there is a little The majority of species outside the tropics rely on a food research work to assess the effect of different factors on resource for their young that become available for a morphometry of testis. Therefore, this study was conducted predictable period each year. The length of the breeding to evaluate the effect of season and daylights on season varies between species, but in the vast majority, morphometry of testis of quail in Bangladesh. the breeding season is asymmetrical with changes in photoperiod. When mature male coturnix were exposed to short photoperiods (6L: 18D) a rapid testicular weight MATERIAL AND METHOD reduction was recorded (Follett & Farner, 1966). Reduction of testicular weight in the common coturnix, exposed to short days (8 h of light), was more rapid in a colder than in Collection of birds. The study was performed on 120 a warmer environment (Wilson et al., 1971). The two most sexually matured domestic quails (Coturnix coturnix critical features in the seasonality of an environment are japonica) weighing average 119.15±7.46 g (range 110 g the amplitude of the seasonal fluctuations and the precision to 130 g). The quail was routinely collected from a with which these fluctuations occur each year (Wingfield recognized farm of Chittagong Metropolitan area, et al., 1992). The physiological processes that underlie the Bangladesh during different seasons of the year. Forty timing of breeding in birds have been studied extensively quails were collected during summer (March-June), 30 * Department of Anatomy and Histology, Faculty of Veterinary Medicine, Chittagong Veterinary & Animal Sciences University, Zakir Hossain Road, Khulshi, Bangladesh. 100 SHIL, S. K.; QUASEM, A. & RAHMAN, M. L. Histological and morphometric analyses of testes of adult quail (Coturnix coturnix japonica) of Bangladesh. Int. J. Morphol., 33(1):100-104, 2015. during rainy (July- September), 20 during autumn RESULTS (October-November) and 30 during winter (December- February). The birds were euthanized (30 mg/kg bw/im) and the testes were collected from each individual bird. Morphometry of testes of quail by season and length of day light. Mean live weight of the Japanese quail (n=120) Gross morphometry. After separation of testis, specimen was 119.15±7.46 g. Average paired testicular weight, volume, was subjected to gross anatomical measurement. Volume length and circumference were significantly higher in summer of testes was measured by Archimedes’ principle of water and rainy seasons than in autumn and winter (P=0.02). The displacement using normal saline (0.85% NaCl) and average weight of testes was significantly heavier (3.17±0.66 results were recorded in cubic milliliters (Bath & g) during long day period (summer and rainy season) than Chaudhari, 2002). Length of testes was measured by that of short day period (1.27±0.28 g) (P=0.03) (Fig. 1). vernier digital calipers (0-150 digital caliper, Shinko Regardless of seasons positive correlation was recorded Denshi Co. Ltd, Japan). Relative testis weight was between the day length and paired testicular weight (r=0.82). measured according to formula as follows: (weight of However, paired testes weight were about 2.5 fold higher in paired testes / body weight) ¥ 100 (Orlu & Egbunike, summer than in winter. Relative testes weight was higher in 2010). summer (2.75±0.50%) and rainy season (2.45±0.44%) than the autumn (1.05±0.22%) and winter (1.13±0.29%). Tissue processing. Sampled testes were immediately immersed in Bouin’s fixative and submitted to prefixation for approximately 20 minutes. Longitudinal sections of testes were then made and sections were immersed in histological fixatives for 24 h in order to complete fixation. Small testis fragments were used for the routine histology such as dehydration, clarification and paraplast embedding and stained with hematoxylin-eosin (HE). Histomorphometry. A total of 12 testes from quails were collected during summer, 9 during rainy, 6 during autumn and 9 during winter season were used for thorough histological study. In order to record histomorphometric Fig. 1. Paired testes weight in relation to natural photoperiod in data 10 photomicrographs were taken randomly from the different seasons. central zone of cross-sections of each testis using a Photomicroscope (AmScope Trinocular compound Morphometry of seminiferous tubule by season and day microscope +1.3 MP camera, Model T490 B-MT) and length. Morphometric evaluation of seminiferous tubules Toup View 3.0 image processing software. Using ImageJ revealed that in summer and rainy season they had their higher 1.45 image measurement software, diameters were diameter, circumference and germinal epithelium height than calculated from 20 randomly selected cross-sections of in autumn and winter (Figs. 2, 3, 4, 5, 6). Thus seminiferous the seminiferous tubules in each testis and recorded their tubules revealed a seasonal variation with two phases: (1) average values. Temperature and day length were recorded reproductive active phase, (2) regressive phase. Elevated from the official website (www.bmd.gov.bd) of values were observed in long day period (May - July) and Bangladesh Meteorological Department of Government decreased progressively until December. A new increase of the People's Republic of Bangladesh. occurred in February, March, followed by an increase in April, May, June and in July it reaches its peak and a progressive Data analysis. Student paired t-tests were used to com- decrease until December. The variation of the height of the pare between paired groups, whereas the one way analysis seminiferous epithelium in general accompanied the annual of variance (ANOVA) was used to compare the differences variation of the tubular diameter. Moderate peaks were in weight, volume, circumference and length of testes, observed in autumn (October, November) and maximum seminiferous tubule diameter, height of germinal values in summer and rainy season i.e. from March to epithelium across the groups. SPSS statistical software September (a marked peak). The lowest values were detected (Ver.16.0 for windows, SPSS) was used for analysis of in winter season (November, December and January). There the data. Results were expressed as arithmetic Mean ± Stan- was strong positive correlation (r=0.84) between dard Deviation (Mean±SD). The differences were seminiferous tubule diameter and paired testes weight considered to be significant if P was ≤0.05. throughout the year. Increase in testes weight also correlated 101 SHIL, S. K.; QUASEM, A. & RAHMAN, M. L. Histological and morphometric analyses of testes of adult quail (Coturnix coturnix japonica) of Bangladesh. Int. J. Morphol., 33(1):100-104, 2015. well with the increase of height (r=0.88) of the germinal epithelium of the seminiferous tubule. Fig. 2. Seasonal variation in histomorphometry of seminiferous tubule. Fig. 3. Histological characters of seminiferous tubule in Summer Fig. 4. Histological characters of seminiferous tubule in Rainy (June) (scale bar 200 µm). season (August) (scale bar 200 µm). Fig. 5. Histological characters of seminiferous tubule in Autumn Fig. 6. Histological characters of seminiferous tubule in Winter (October) (scale bar 200 mm) (January) (scale bar 200 mm). 102 SHIL, S. K.; QUASEM, A. & RAHMAN, M. L. Histological and morphometric analyses of testes of adult quail (Coturnix coturnix japonica) of Bangladesh. Int. J. Morphol., 33(1):100-104, 2015.
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